PH DEPENDENT PHOTOPHYSICS OF PHLUORIN GFP MUTANTS

这个项目的总体目标是了解细胞 神经调节剂控制行为可塑性的机制。 具体地说，我们正在使用多光子显微镜和 电生理学用来研究细胞机制的 神经调节剂在一个小井中产生各种运动模式 特色化的神经网络--龙虾的口胃神经节。 神经调节剂，如多巴胺和5-羟色胺，在 脊椎动物和无脊椎动物行为的调节。这个 哈里斯-沃里克研究小组分析了多巴胺、5-羟色胺 和章鱼胺在幽门14细胞网络上的作用 甲壳类口胃神经节(STG)。这些化合物是 幽门网络的内源性神经调节剂及何时 沐浴--应用于口胃神经节，每种胺都能激起一种 幽门产生的节律运动模式的独特变异 网络，随着周期频率的变化，活跃的神经元，相位 以及神经元激发的强度。我们目前使用的是多光子 显微镜测量钙信号并监测时间进程和 大鼠STG神经元钙离子变化的空间分布 胺。这些实验旨在回答两个主要问题： (1)胺是否影响细胞内钙的进入和/或释放 储存，和(2)是胺对钙活性的作用 神经末梢不同(它们可能影响递质 释放)，而不是在细胞的其他区域，其中钙可能 调节细胞的固有激发特性？

The overall goal of this project is to understand the cellular mechanisms used by neuromodulators to control behavioural plasticity. Specifically, we are using multiphoton microscopy and electrophysiology to study the cellular mechanisms by which neuromodulators produce various motor patterns in a small, well characterized, neural network-- the lobster stomatogastric ganglion. Neuromodulators such as dopamine and serotonin play central roles in the regulation of behavior in both vertebrates and invertebrates. The Harris-Warrick group has analyzed the effects of dopamine, serotonin and octopamine on the well-studied 14-cell pyloric network in the crustacean stomatogastric ganglion (STG). These compounds are endogenous neuromodulators of the pyloric network and when bath-applied to the stomatogastric ganglion, each amine evokes a unique variant on the rhythmic motor pattern generated by the pyloric network, with changes in the cycle frequency, active neurons, phasing and intensity of n euronal firing. We are currently using multiphoton microscopy measure of calcium signals and monitor the time course and spatial distribution of the calcium changes in STG neurons elicited by amines. These experiments are designed to answer two major questions: (1) do amines modify calcium entry and/or release from intracellular stores, and (2) are the actions of amines on calcium activity different at nerve terminals (where they might affect transmitter release) than in other regions of the cell where calcium might modulate the intrinsic firing properties of the cell?